Search results for: fluorescence polarization

Authors: Endris Yibru Hanurry, Wei-Hsin Hsu, Hsieh-Chih Tsai

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In recent years, the discovery of small interfering RNAs (siRNAs) has got great attention for the treatment of cancer and other diseases. However, the therapeutic efficacy of siRNAs has been faced with many drawbacks because of short half-life in blood circulation, poor membrane penetration, weak endosomal escape and inadequate release into the cytosol. To overcome these drawbacks, we designed a non-viral vector by conjugating polyamidoamine generation 4.5 dendrimer (PDG4.5) with diethylenetriamine (DETA)- and tetraethylenepentamine (TEPA) followed by binding with siRNA to form polyplexes through electrostatic interaction. The result of 1H nuclear magnetic resonance (NMR), 13C NMR, correlation spectroscopy, heteronuclear single–quantum correlation spectroscopy, and Fourier transform infrared spectroscopy confirmed the successful conjugation of DETA and TEPA with PDG4.5. Then, the size, surface charge, morphology, binding ability, stability, release assay, toxicity and cellular internalization were analyzed to explore the physicochemical and biological properties of PDG4.5-DETA and PDG4.5-TEPA polyplexes at specific N/P ratios. The polyplexes (N/P = 8) exhibited spherical nanosized (125 and 85 nm) particles with optimum surface charge (13 and 26 mV), showed strong siRNA binding ability, protected the siRNA against enzyme digestion and accepted biocompatibility to the HeLa cells. Qualitatively, the fluorescence microscopy image revealed the delocalization (Manders’ coefficient 0.63 and 0.53 for PDG4.5-DETA and PDG4.5-TEPA, respectively) of polyplexes and the translocation of the siRNA throughout the cytosol to show a decent cellular internalization and intracellular biodistribution of polyplexes in HeLa cells. Quantitatively, the flow cytometry result indicated that a significant (P < 0.05) amount of siRNA was internalized by cells treated with PDG4.5-DETA (68.5%) and PDG4.5-TEPA (73%) polyplexes. Generally, PDG4.5-DETA and PDG4.5-TEPA were ideal nanocarriers of siRNA in vitro and might be used as promising candidates for in vivo study and future pharmaceutical applications.

Keywords: non-viral carrier, oligoalkylamine, poly(amidoamine) dendrimer, polyplexes, siRNA

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Authors: Fei Ye, Åsa Barrefelt, Manuchehr Abedi-Valugerdi, Khalid M. Abu-Salah, Salman A. Alrokayan, Mamoun Muhammed, Moustapha Hassan

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With appropriate encapsulation in functional nanoparticles drugs are more stable in physiological environment and the kinetics of the drug can be more carefully controlled and monitored. Furthermore, targeted drug delivery can be developed to improve chemotherapy in cancer treatment, not only by enhancing intracellular uptake by target cells but also by reducing the adverse effects in non-target organs. Inorganic imaging agents, delivered together with anti-cancer drugs, enhance the local imaging contrast and provide precise diagnosis as well as evaluation of therapy efficacy. We have developed biodegradable polymeric vesicles as a nanocarrier system for multimodal bio-imaging and anticancer drug delivery. The poly (lactic-co-glycolic acid) PLGA) vesicles were fabricated by encapsulating inorganic imaging agents of superparamagnetic iron oxide nanoparticles (SPION), manganese-doped zinc sulfide (MN:ZnS) quantum dots (QDs) and the anticancer drug busulfan into PLGA nanoparticles via an emulsion-evaporation method. T2-weighted magnetic resonance imaging (MRI) of PLGA-SPION-Mn:ZnS phantoms exhibited enhanced negative contrast with r2 relaxivity of approximately 523 s-1 mM-1 Fe. Murine macrophage (J774A) cellular uptake of PLGA vesicles started fluorescence imaging at 2 h and reached maximum intensity at 24 h incubation. The drug delivery ability PLGA vesicles was demonstrated in vitro by release of busulfan. PLGA vesicles degradation was studied in vitro, showing that approximately 32% was degraded into lactic and glycolic acid over a period of 5 weeks. The biodistribution of PLGA vesicles was investigated in vivo by MRI in a rat model. Change of contrast in the liver could be visualized by MRI after 7 min and maximal signal loss detected after 4 h post-injection of PLGA vesicles. Histological studies showed that the presence of PLGA vesicles in organs was shifted from the lungs to the liver and spleen over time.

Keywords: biodegradable polymers, multifunctional nanoparticles, quantum dots, anticancer drugs

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Authors: Sospeter P. Maganga, Alphonce Wikedzi, Mussa D. Budeba, Samwel V. Manyele

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This study characterizes complex gold ores (elemental and mineralogical composition, gold distribution, ore grindability, and mineral liberation) for preliminary process selection. About 200 kg of ore samples were collected from each location using systematic sampling by mass interval. Ores were dried, crushed, milled, and split into representative sub-samples (about 1 kg) for elemental and mineralogical composition analyses using X-ray fluorescence (XRF), fire assay finished with Atomic Absorption Spectrometer (AAS), and X-ray Diffraction (XRD) methods, respectively. The gold distribution was studied on size-by-size fractions, while ore grindability was determined using the standard Bond test. The mineral liberation analysis was conducted using ThermoFisher Scientific Mineral Liberation Analyzer (MLA) 650, where unsieved polished grain mounts (80% passing 700 µm) were used as MLA feed. Two MLA measurement modes, X-ray modal analysis (XMOD) and sparse phase liberation-grain X-ray mapping analysis (SPL-GXMAP), were employed. At least two cyanide consumers (Cu, Fe, Pb, and Zn) and kinetics impeders (Mn, S, As, and Bi) were present in all locations investigated. Copper content at Kapanda (0.77% Cu) and Ibindi (7.48% Cu) exceeded the recommended threshold of 0.5% Cu for direct cyanidation. The gold ore at Ibindi indicated a higher rate of grinding compared to other locations. This could be explained by the highest grindability (2.119 g/rev.) and lowest Bond work index (10.213 kWh/t) values. The pyrite-marcasite, chalcopyrite, galena, and siderite were identified as major gold, copper, lead, and iron-bearing minerals, respectively, with potential for economic extraction. However, only gold and copper can be recovered under conventional milling because of grain size issues (galena is exposed by 10%) and process complexity (difficult to concentrate and smelt iron from siderite). Therefore, the preliminary process selection is copper flotation followed by gold cyanidation for Kapanda and Ibindi ores, whereas gold cyanidation with additives such as glycine or ammonia is selected for Mawemeru and Itumbi ores because of low concentrations of Cu, Pb, Fe, and Zn minerals.

Keywords: complex gold ores, mineral liberation, ore characterization, ore grindability

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Authors: Feixiong Chen, Naoufel Haddour, Marie Frenea-Robin, Yves MéRieux, Yann Chevolot, Virginie Monnier

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Neonatal thrombopenia occurs when the mother generates antibodies against her baby’s platelet antigens. It is particularly critical for newborns because it can cause coagulation troubles leading to intracranial hemorrhage. In this case, diagnosis must be done quickly to make platelets transfusion immediately after birth. Before transfusion, platelet antigens must be tested carefully to avoid rejection. The majority of thrombopenia (95 %) are caused by antibodies directed against Human Platelet Antigen 1a (HPA-1a) or 5b (HPA-5b). The common method for antigen platelets detection is polymerase chain reaction allowing for identification of gene sequence. However, it is expensive, time-consuming and requires significant blood volume which is not suitable for newborns. We propose to develop a point-of-care device based on double functionalized magnetic colloids with 1) antibodies specific to antigen platelets and 2) highly sensitive electroactive molecules in order to be detected by an electrochemical microsensor. These magnetic colloids will be used first to isolate platelets from other blood components, then to capture specifically platelets bearing HPA-1a and HPA-5b antigens and finally to attract them close to sensor working electrode for improved electrochemical signal. The expected advantages are an assay time lower than 20 min starting from blood volume smaller than 100 µL. Our functionalization procedure based on amine dendrimers and NHS-ester modification of initial carboxyl colloids will be presented. Functionalization efficiency was evaluated by colorimetric titration of surface chemical groups, zeta potential measurements, infrared spectroscopy, fluorescence scanning and cyclic voltammetry. Our results showed that electroactive molecules and antibodies can be immobilized successfully onto magnetic colloids. Application of a magnetic field onto working electrode increased the detected electrochemical signal. Magnetic colloids were able to capture specific purified antigens extracted from platelets.

Keywords: Magnetic Nanoparticles , Electroactive Molecules, Antibody, Platelet

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Authors: Eva Slaninova, Diana Cernayova, Zuzana Sedrlova, Katerina Mrazova, Petr Sedlacek, Jana Nebesarova, Stanislav Obruca

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Cyanobacteria are gram-negative prokaryotes belonging to a group of photosynthetic bacteria. In comparison with heterotrophic microorganisms, cyanobacteria utilize atmospheric nitrogen and carbon dioxide without any additional substrates. This ability of these microorganisms could be employed in biotechnology for the production of bioplastics, concretely polyhydroxyalkanoates (PHAs) which are primarily accumulated as a storage material in cells in the form of intracellular granules. In this study, there two cyanobacterial cultures from genera Synechocystis were used, namely Synechocystic sp. PCC 6803 and Synechocystis salina CCALA 192. There were optimized and used several various approaches, including microscopic techniques such as cryo-scanning electron microscopy (Cryo-SEM) and transmission electron microscopy (TEM), and fluorescence lifetime imaging microscopy using Nile red as a fluorescent probe (FLIM). Due to these instrumental techniques, the morphology of intracellular space and surface of cells were characterized. The next group of methods which were employed was spectroscopic techniques such as UV-Vis spectroscopy measured in two modes (turbidimetry and integration sphere) and Fourier transform infrared spectroscopy (FTIR). All these diverse techniques were used for the detection and characterization of pigments (chlorophylls, carotenoids, phycocyanin, etc.) and PHAs, in our case poly (3-hydroxybutyrate) (P3HB). To verify results, gas chromatography (GC) was employed concretely for the determination of the amount of P3HB in biomass. Cyanobacteria were also characterized as polyhydroxybutyrate producers by flow cytometer, which could count cells and at the same time distinguish cells including P3HB and without due to fluorescent probe called BODIPY and live/dead fluorescent probe SYTO Blue. Based on results, P3HB content in cyanobacteria cells was determined, as also the overall fitness of the cells. Acknowledgment: Funding: This study was partly funded by the projectGA19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), project I 4082-B25.

Keywords: cyanobacteria, fluorescent probe, microscopic techniques, poly(3hydroxybutyrate), spectroscopy, chromatography

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Authors: Patarasuda Chaisupa, R. Clay Wright

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The selection of appropriate biomolecular targets is a crucial aspect of biopharmaceutical development. The Auxin-Inducible Degron Degradation (AID) technology has demonstrated remarkable potential in efficiently and rapidly degrading target proteins, thereby enabling the identification and acquisition of drug targets. The AID system also offers a viable method to deplete specific proteins, particularly in cases where the degradation pathway has not been exploited or when the adaptation of proteins, including the cell environment, occurs to compensate for the mutation or gene knockout. In this study, we have engineered an improved AID system tailored to deplete proteins of interest. This AID construct combines the auxin-responsive E3 ubiquitin ligase binding domain, AFB2, and the substrate degron, IAA17, fused to the target genes. Essential genes of fungi with the lowest percent amino acid similarity to human and plant orthologs, according to the Basic Local Alignment Search Tool (BLAST), were cloned into the AID construct in S. cerevisiae (AID-tagged strains) using a modular yeast cloning toolkit for multipart assembly and direct genetic modification. Each E3 ubiquitin ligase and IAA17 degron was fused to a fluorescence protein, allowing for real-time monitoring of protein levels in response to different auxin doses via cytometry. Our AID system exhibited high sensitivity, with an EC50 value of 0.040 µM (SE = 0.016) for AFB2, enabling the specific promotion of IAA17::target protein degradation. Furthermore, we demonstrate how this improved AID system enhances quantitative functional studies of various proteins in fungi. The advancements made in auxin-inducible protein degradation in this study offer a powerful approach to investigating critical target protein viability in fungi, screening protein targets for drugs, and regulating intracellular protein abundance, thus revolutionizing the study of protein function underlying a diverse range of biological processes.

Keywords: synthetic biology, bioengineering, molecular biology, biotechnology

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Authors: J. Oustric, L. Berti, J. Santini

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Nowadays, the objective of durable agriculture, and in particular organic agriculture, is to reduce the level of fertilizer inputs used in crops. Limiting the quantity of fertilizer inputs would optimize the economical result and minimizing the environmental impact. Nutrient deficiency, particularly of a major nutrient (N, P, and K), can seriously affect fruit production and quality. In citrus crops, rootstock/scion combinations. In citrus crop, scion/rootstock combinations are used frequently to improve tolerance to various abiotic stresses. New rootstocks are needed to respond to these constraints, and the use of new tetraploid rootstocks better adapted to lower nutrient intake could offer a promising way forward. The aim of this work was to determine whether a better tolerance to nutrient deficiency could be observed in a doubled diploid seedling and whether this tolerance could be observed in common clementine scion if used as rootstocks. We selected diploid (CM2x) and doubled diploid (CM4x) Citrumelo 4475 seedlings and common clementine (C) grafted onto Citrumelo 4475 diploid (C/CM2x) and doubled diploid (C/CM4x) rootstocks. Nutrient deficiency effects on the seedlings and scion/rootstock combinations were analyzed by studying anatomical, structural and ultrastructural determinants (chlorosis, stomata, ostiole and cells and their organelles), photosynthetic properties (leaf net photosynthetic rate (Pₙₑₜ), stomatal conductance (gₛ), chlorophyll a fluorescence (Fᵥ/Fₘ)) and oxidative marker (malondialdehyde). Nutrient deficiency affected differently foliar tissues, physiological parameters, and oxidative metabolism in leaves of seedlings depending on their ploidy level and of common clementine scion depending on their rootstocks ploidy level. Both CM4x and C/CM4x presented lower foliar damages (chlorosis, chloroplasts, mitochondria, and plastoglobuli), photosynthesis processes alteration (Pₙₑₜ, gₛ, and Fᵥ/Fₘ), and malondialdehyde accumulation than CM2x and C/CM2x after nutrient deficiency. Doubled diploid Citrumelo 4475 can improve nutrient deficiency tolerance, and its use as a rootstock allows to confer this tolerance to the common clementine scion.

Keywords: nutrient deficiency, oxidative stress, photosynthesis, polyploid rootstocks

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Authors: Khairil Ahmad, Jenny Gryzelius, Mohd Helmi Mohd Sobri

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With the waning support for centrist ‘third-way’ politics across the Western world, there has been an increase in political parties and individual candidates relying on populist political discourse and rhetoric in order to capitalize on the sense of frustration apparent within the electorate. What is of note is the divergence in the discourses employed. On the one hand, there is a polarization between a growing wave of populist right-wing parties and politicians, employing a mixture of economic populism with divisive nationalistic ideals such as restricted immigration, for example, the UK’s UKIP and Donald Trump in the US. On the other hand, there are resurgent, often grassroots-led, left-wing movements and politicians, such as Podemos in Spain and Jeremy Corbyn in the UK, focusing on anti-austerity measures and inclusive policies. In general, the concept of populism is often ascribed in a pejorative way. This is despite the success of populist left-wing governments across Latin America in recent times, especially in terms of reducing poverty. Nonetheless, recently, scholars such as Ernesto Laclau have tried to rethink populism as a social scientific concept which is essential in helping us make sense of contemporary political articulations. Using Laclau’s framework, this paper seeks to analyze poverty reduction policies in different iterations in the context of the tenures of three Prime Ministers of Malaysia. The first is Abdul Razak Hussein’s New Economic Policy, which focused on uplifting the economic position of Malaysia’s majority Malay population. The second is Mahathir Mohamad’s state-led neo-liberalization of the Malaysian economy, which focused on the creation of a core group of crony elites in order to spearhead economic development. The third is current Prime Minister Najib Razak’s targeted poverty eradication strategy through a focused program which directly provides benefits to recipients such as through direct cash transfers. The paper employs a discursive approach to trace elements of populism in these cases and highlight instances of how their strategies are articulated in ways that seek to appeal towards particular visions of national unity.

Keywords: discourse analysis, Malaysia, populism, poverty eradication

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Authors: Ali Bahkali, Rayan Yousef Booq, Mohammad Khiyami

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Soil samples were collected from five different regions in the Kingdom of Saudi Arabia. Microbiological methods included dilution methods and pour plates to isolate and purify bacteria soil. The ability of isolates to develop biopolymer was investigated on petri dishes containing elements and substance concentrations stimulating developing biopolymer. Fluorescent stains, Nile red and Nile blue were used to stain the bacterial cells developing biopolymers. In addition, Sudan black was used to detect biopolymers in bacterial cells. The isolates which developed biopolymers were identified based on their gene sequence of 1 6sRNA and their ability to grow and synthesize PHAs on mineral medium supplemented with 1% dates molasses as the only carbon source under nitrogen limitation. During the study 293 bacterial isolates were isolated and detected. Through the initial survey on the petri dishes, 84 isolates showed the ability to develop biopolymers. These bacterial colonies developed a pink color due to accumulation of the biopolymers in the cells. Twenty-three isolates were able to grow on dates molasses, three strains of which showed the ability to accumulate biopolymers. These strains included Bacillus sp., Ralstonia sp. and Microbacterium sp. They were detected by Nile blue A stain with fluorescence microscopy (OLYMPUS IX 51). Among the isolated strains Ralstonia sp. was selected after its ability to grow on molasses dates in the presence of a limited nitrogen source was detected. The optimum conditions for formation of biopolymers by isolated strains were investigated. Conditions studied included, best incubation duration (2 days), temperature (30°C) and pH (7-8). The maximum PHB production was raised by 1% (v1v) when using concentrations of dates molasses 1, 2, 3, 4 and 5% in MSM. The best inoculated with 1% old inoculum (1= OD). The ideal extraction method of PHA and PHB proved to be 0.4% sodium hypochlorite solution, producing a quantity of polymer 98.79% of the cell's dry weight. The maximum PHB production was 1.79 g/L recorded by Ralstonia sp. after 48 h, while it was 1.40 g/L produced by R.eutropha ATCC 17697 after 48 h.

Keywords: bacteria forming polyhydroxyalkanoate, detection, molecular, Saudi Arabia

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Authors: Mohammad Nasb, Muhammad Rehan, Chen Hong

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Background Meniscus defects critically alter knee function and lead to degenerative changes. Regenerative medicine applications including stem cell transplantation have showed a promising efficacy in finding alternatives to overcome traditional treatment limitations. However, stem cell therapy remains limited due to the substantially reduced viability and inhibitory microenvironment. Since tissue growth and repair are under the control of biochemical and mechanical signals, several approaches have recently been investigated (e.g., low intensity pulsed ultrasound [LIPUS]) to promote the regeneration process. This study employed LIPUS to improve growth and osteogenic differentiation of mesenchymal stem cells derived from human embryonic stem cells to improve the regeneration of meniscus tissue. Methodology: The Mesenchymal stromal cells (MSCs) were transplanted into the epicenter of the injured meniscus in rabbits, which were randomized into two main groups: a treatment group (n=32 New Zealand rabbits) including 4 subgroups of 8 rabbits in each subgroup (LIPUS treatment, MSC treatment, LIPUS with MSC and control), and a second group (n=9) to track implanted cells and their progeny using green fluorescence protein (GFP). GFP consists of the MSC and LIPUS-MSC combination subgroups. Rabbits were then subjected to histological, immunohistochemistry, and MRI assessment. Results: The quantity of the newly regenerated tissue in the combination treatment group that had Ultrasound irradiation after mesenchymal stem cells were better at all end points. Likewise, Tissue quality scores were also greater in knees treated with both approaches compared with controls and single treatment at all end points, achieving significance at twelve and twenty-four weeks [p < 0.05], and [p = 0.008] at twelve weeks. Differentiation into type-I and II collagen-expressing cells were higher in the combination group at up to twenty-four weeks. Conclusions: the combination of mesenchymal stem cells and LIPUS showed greater adhering to the sites of meniscus injury, differentiate into cells resembling meniscal fibrochondrocytes, and improve both quality and quantity of meniscal regeneration.

Keywords: stem cells, regenerative medicine, osteoarthritis, knee

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Authors: Chabungbam Niranjit Khuman, Makarand Madhao Ghangrekar, Arunabha Mitra

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The cost of aeration is one of the limiting factors in the upscaling of microbial fuel cells (MFC) for field-scale applications. Wetland macrophytes have the ability to release oxygen into the water to maintain aerobic conditions in their root zone. In this experiment, the efficacy of rhizospheric oxygen release of wetland macrophytes as a source of oxygen in the cathodic chamber of MFC was conducted. The experiment was conducted in an MFC consisting of a three-liter anodic chamber made of ceramic cylinder and a 27 L cathodic chamber. Untreated carbon felts were used as electrodes (i.e., anode and cathode) and connected to an external load of 100 Ω using stainless steel wire. Wetland macrophytes (Canna indica) were grown in the cathodic chamber of the MFC in a hydroponic fashion using a styrofoam sheet (termed as macrophytes assisted-microbial fuel cell, M-MFC). The catholyte (i.e., water) in the M-MFC had negligible contact with atmospheric air due to the styrofoam sheet used for maintaining the hydroponic condition. There was no mixing of the catholyte in the M-MFC. Sucrose based synthetic wastewater having chemical oxygen demand (COD) of 3000 mg/L was fed into the anodic chamber of the MFC in fed-batch mode with a liquid retention time of four days. The C. indica thrived well throughout the duration of the experiment without much care. The average dissolved oxygen (DO) concentration and pH value in the M-MFC were 3.25 mg/L and 7.07, respectively, in the catholyte. Since the catholyte was not in contact with air, the DO in the catholyte might be considered as solely liberated from the rhizospheric oxygen release of C. indica. The maximum COD removal efficiency of M-MFC observed during the experiment was 76.9%. The inadequacy of terminal electron acceptor in the cathodic chamber in M-MFC might have hampered the electron transfer, which in turn, led to slower specific microbial activity, thereby resulting in lower COD removal efficiency than the traditional MFC with aerated catholyte. The average operating voltage (OV) and open-circuit voltage (OCV) of 294 mV and 594 mV, respectively, were observed in M-MFC. The maximum power density observed during polarization was 381 mW/m³, and the maximum sustainable power density observed during the experiment was 397 mW/m³ in M-MFC. The maximum normalized energy recovery and coulombic efficiency of 38.09 Wh/m³ and 1.27%, respectively, were observed. Therefore, it was evidenced that rhizospheric oxygen release of wetland macrophytes (C. indica) was capable of sustaining the cathodic reaction in MFC for field-scale applications.

Keywords: hydroponic, microbial fuel cell, rhizospheric oxygen release, wetland macrophytes

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Authors: G. Amariei, K. Boltes, R. Rosal, P. Leton

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Wastewater treatment plants (WWTPs) are supposed to hold an important place in the reduction of emerging contaminants, but provide an environment that has potential for the development and/or spread of adaptation, as bacteria are continuously mixed with contaminants at sub-inhibitory concentrations. Reviewing the literature, there are little data available regarding the use of adapted bacteria forming activated sludge community for toxicity assessment, and only individual validations have been performed. Therefore, the aim of this work was to study the toxicity of Triclosan (TCS) and Ibuprofen (IBU), individually and in binary combination, on adapted activated sludge (AS). For this purpose a battery of biomarkers were assessed, involving oxidative stress and cytotoxicity responses: glutation-S-transferase (GST), catalase (CAT) and viable cells with FDA. In addition, we compared the toxic effects on adapted bacteria with unadapted bacteria, from a previous research. Adapted AS comes from three continuous-flow AS laboratory systems; two systems received IBU and TCS, individually; while the other received the binary combination, for 14 days. After adaptation, each bacterial culture condition was exposure to IBU, TCS and the combination, at 12 h. The concentration of IBU and TCS ranged 0.5-4mg/L and 0.012-0.1 mg/L, respectively. Batch toxicity experiments were performed using Oxygraph system (Hansatech), for determining the activity of CAT enzyme based on the quantification of oxygen production rate. Fluorimetric technique was applied as well, using a Fluoroskan Ascent Fl (Thermo) for determining the activity of GST enzyme, using monochlorobimane-GSH as substrate, and to the estimation of viable cell of the sludge, by fluorescence staining using Fluorescein Diacetate (FDA). For IBU adapted sludge, CAT activity it was increased at low concentration of IBU, TCS and mixture. However, increasing the concentration the behavior was different: while IBU tends to stabilize the CAT activity, TCS and the mixture decreased this one. GST activity was significantly increased by TCS and mixture. For IBU, no variations it was observed. For TCS adapted sludge, no significant variations on CAT activity it was observed. GST activity it was significant decreased for all contaminants. For mixture adapted sludge the behaviour of CAT activity it was similar to IBU adapted sludge. GST activity it was decreased at all concentration of IBU. While the presence of TCS and mixture, respectively, increased the GST activity. These findings were consistent with the viability cells evaluation, which clearly showed a variation of sludge viability. Our results suggest that, compared with unadapted bacteria, the adapted bacteria conditions plays a relevant role in the toxicity behaviour towards activated sludge communities.

Keywords: adapted sludge community, mixture, PPCPs, toxicity

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Authors: Gulnaz Khan, Meha F. Aftab, Munazza Murtaza, Rizwana S. Waraich

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Advanced glycation end products (AGEs) precursor and its abnormal accumulation cause damage to various tissues and organs. AGEs have pathogenic implication in several diseases including diabetes. Existing AGEs inhibitors are not in clinical use, and there is a need for development of novel inhibitors. The present investigation aimed at identifying the novel AGEs inhibitors and assessing their mechanism of action for treating insulin resistance in mice model of diabetes. Novel derivatives of benzylidene of indan-1,3-dione were synthesized. The compounds were selected to study their action mechanism in improving insulin resistance, in vitro, in human hepatocytes and murine adipocytes and then, in vivo, in mice genetic model of diabetes (db/db). Mice were treated with novel derivatives of benzylidene of indane 1,3-dione. AGEs mediated ROS production was measured by dihydroethidium fluorescence assay. AGEs level in the serum of treated mice was observed by ELISA. Gene expression of receptor for AGEs (RAGE), PPAR-gamma, TNF-alpha and GLUT-4 was evaluated by RT-PCR. Glucose uptake was measured by fluorescent method. Microscopy was used to analyze glycogen synthesis in muscle. Among several derivatives of benzylidene of indan-1,3-dione, IDD-24, demonstrated highest inhibition of AGESs. IDD-24 significantly reduced AGEs formation and expression of receptor for advanced glycation end products (RAGE) in fat, liver of db/db mice. Suppression of AGEs mediated ROS production was also observed in hepatocytes and fat cell, after treatment with IDD-24. Glycogen synthesis was increased in muscle tissue of mice treated with IDD-24. In adipocytes, IDD-24 prevented AGEs induced reduced glucose uptake. Mice treated with IDD-24 exhibited increased glucose tolerance, serum adiponectin levels and decreased insulin resistance. The result of present study suggested that IDD-24 can be a possible treatment target to address glycotoxins induced insulin resistance.

Keywords: advance glycation end product, hyperglycemia, indan-1, 3-dione, insulin resistance

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Authors: Recep Keşli, Hayriye Tokay, Cengiz Demir, İsmail Ceyhan

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Objective: In many countries, tuberculosis (TB) is still one of the most important diseases. Tuberculosis is among top 10 causes of death worldwide. The early diagnosis of active tuberculosis still depends on the presence of acid resistant bacilli (ARB) in stained smears. In this study, we aimed to investigate the diagnostic performances of Erlich Ziehl Neelsen (EZN), Kinyoun and two different fluorescent stains. Methods: The specimens were obtained from the patients who applied to Chest Diseases Departments of Ankara Atatürk Chest Diseases and Thoracic Surgery Training and Research Hospital, and Afyon Kocatepe University, ANS Research and Practice Hospital. The study was carried out in the Medical Microbiology Laboratory, School of Medicine, Afyon Kocatepe University. All the non-sterile specimens were homogenized and decontaminated according to the EUCAST instructions. Samples were inoculated onto the Löwenstein-Jensen agars (bio-Merieux Marcy l'Etoile, France) and then incubated at 37˚C, for 40 days. Four smears were prepared from each specimen. Slides were stained with commercial EZN (BD, Sparks, USA), Kinyoun (SALUBRIS Istanbul, Turkey), Auramine (SALUBRIS Istanbul, Turkey) and Rhodamine (SALUBRIS Istanbul, Turkey) kit. While EZN and Kinyoun stainings were examined by light microscope, Auramine and Rhodamine slides were examined by fluorescence microscopy. Results: A total of 158 respiratory system samples (sputum, broncho alveolar lavage fluid…etc) were enrolled into the study. A hundred and two of the samples that processed were found as culture positive. The sensitivity, specificity, positive predictive, and negative predictive values were detected as 100%, 67.5%, 73.5%, and 100% for EZN, 100%, 70.9%, 77.4%, and 100% for Kinyoun, 100%,77.8%, 84.3%, 100% for Auramine, and 100%, 80% , 86.3%, and 100% for Rhodamine respectively. Conclusions: According to our study auramine and rhodamine staining methods showed the best diagnostic performance among the four investigated staining methods. In conclusion, the fluorochrome staining method may be accepted as the most reliable, rapid and useful method for diagnosis of the mycobacterial infections truly.

Keywords: acid resistant bacilli (ARB), auramine, Ehrlich-Ziehl-Neelsen (EZN), Kinyoun, Rhodamine

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Authors: Amanda J. Burridge, Keith J. Edwards, Paul A. Wilkinson, Tom Batstone, Erik H. Murchie, Lorna McAusland, Ana Elizabete Carmo-Silva, Ivan Jauregui, Tracy Lawson, Silvere R. M. Vialet-Chabrand

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The rate of genetic progress in wheat production must be improved to meet global food security targets. However, past selection for domestication traits has reduced the genetic variation in modern wheat cultivars, a fact that could severely limit the future rate of genetic gain. The genetic variation in agronomically important traits for the wild relatives and progenitors of wheat is far greater than that of the current domesticated cultivars, but transferring these traits into modern cultivars is not straightforward. Between the elite cultivars of wheat, photosynthetic capacity is a key trait for which there is limited variation. Early screening of wheat wild relative and progenitors has shown differences in photosynthetic capacity and efficiency not only between wild relative species but marked differences between the accessions of each species. By identifying wild relative accessions with improved photosynthetic traits and characterising the genetic variation responsible, it is possible to incorporate these traits into advanced breeding programmes by wide crossing and introgression programmes. To identify the potential variety of photosynthetic capacity and efficiency available in the secondary and tertiary genepool, a wide scale survey was carried out for over 600 accessions from 80 species including those from the genus Aegilops, Triticum, Thinopyrum, Elymus, and Secale. Genotype data were generated for each accession using a ‘Wheat Wild Relative’ Single Nucleotide Polymorphism (SNP) genotyping array composed of 35,000 SNP markers polymorphic between wild relatives and elite hexaploid wheat. This genotype data was combined with phenotypic measurements such as gas exchange (CO₂, H₂O), chlorophyll fluorescence, growth, morphology, and RuBisCO activity to identify potential breeding material with enhanced photosynthetic capacity and efficiency. The data and associated analysis tools presented here will prove useful to anyone interested in increasing the genetic diversity in hexaploid wheat or the application of complex genotyping data to plant breeding.

Keywords: wheat, wild relatives, pre-breeding, genomics, photosynthesis

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Authors: Christoph Lohr, Hanna Wund, Peter Elsner, Kay André Weidenmann

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Injection molding is one of the most commonly used techniques in the industrial polymer processing. In the conventional process of injection molding, the liquid polymer is injected into the cavity of the mold, where the polymer directly starts hardening at the cooled walls. To compensate the shrinkage, which is caused predominantly by the immediate cooling, holding pressure is applied. Through that whole process, residual stresses are produced by the temperature difference of the polymer melt and the injection mold and the relocation of the polymer chains, which were oriented by the high process pressures and injection speeds. These residual stresses often weaken or change the structural behavior of the parts or lead to deformation of components. One solution to reduce the residual stresses is the use of variothermal processing. Hereby the mold is heated – i.e. near/over the glass transition temperature of the polymer – the polymer is injected and before opening the mold and ejecting the part the mold is cooled. For the next cycle, the mold gets heated again and the procedure repeats. The rapid heating and cooling of the mold are realized indirectly by convection of heated and cooled liquid (here: water) which is pumped through fluid channels underneath the mold surface. In this paper, the influences of variothermal processing on the residual stresses are analyzed with samples in a larger scale (500 mm x 250 mm x 4 mm). In addition, the influence on functional elements, such as abrupt changes in wall thickness, bosses, and ribs, on the residual stress is examined. Therefore the polycarbonate samples are produced by variothermal and isothermal processing. The melt is injected into a heated mold, which has in our case a temperature varying between 70 °C and 160 °C. After the filling of the cavity, the closed mold is cooled down varying from 70 °C to 100 °C. The pressure and temperature inside the mold are monitored and evaluated with cavity sensors. The residual stresses of the produced samples are illustrated by birefringence where the effect on the refractive index on the polymer under stress is used. The colorful spectrum can be uncovered by placing the sample between a polarized light source and a second polarization filter. To show the achievement and processing effects on the reduction of residual stress the birefringence images of the isothermal and variothermal produced samples are compared and evaluated. In this comparison to the variothermal produced samples have a lower amount of maxima of each color spectrum than the isothermal produced samples, which concludes that the residual stress of the variothermal produced samples is lower.

Keywords: birefringence, injection molding, polycarbonate, residual stress, variothermal processing

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Authors: Mohamed Moustafa, Medhat Abdallah, Ramy Magdy, Ahmed Abdrabou, Mohamed Badr

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The scribe box under study dates back to the old kingdom. It was excavated by the Italian expedition in Qena (1935-1937). The box consists of 2pieces, the lid and the body. The inner side of the lid is decorated with ancient Egyptian inscriptions written with a black pigment. The box was made using several panels assembled together by wooden dowels and secured with plant ropes. The entire box is covered with a red pigment. This study aims to use analytical techniques in order to identify and have deep understanding for the box components. Moreover, the authors were significantly interested in using infrared reflectance transmission imaging (RTI-IR) to improve the hidden inscriptions on the lid. The identification of wood species included in this study. The visual observation and assessment were done to understand the condition of this box. 3Ddimensions and 2D programs were used to illustrate wood joints techniques. Optical microscopy (OM), X-ray diffraction (XRD), X-ray fluorescence portable (XRF) and Fourier Transform Infrared spectroscopy (FTIR) were used in this study in order to identify wood species, remains of insects bodies, red pigment, fibers plant and previous conservation adhesives, also RTI-IR technique was very effective to improve hidden inscriptions. The analysis results proved that wooden panels and dowels were identified as Acacia nilotica, wooden rail was Salix sp. the insects were identified as Lasioderma serricorne and Gibbium psylloids, the red pigment was Hematite, while the fiber plants were linen, previous adhesive was identified as cellulose nitrates. The historical study for the inscriptions proved that it’s a Hieratic writings of a funerary Text. After its transportation from the Egyptian museum storage to the wood conservation laboratory of the Grand Egyptian museum –conservation center (GEM-CC), conservation techniques were applied with high accuracy in order to restore the object including cleaning , consolidating of friable pigments and writings, removal of previous adhesive and reassembly, finally the conservation process that were applied were extremely effective for this box which became ready for display or storage in the grand Egyptian museum.

Keywords: scribe box, hieratic, 3D program, Acacia nilotica, XRD, cellulose nitrate, conservation

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Authors: Mohamed H. Sorour, Hayam F. Shaalan, Heba A. Hani, Eman S. Sayed, Amany A. El-Mansoup

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Heavy metals (HMs) present an increasing threat to aquatic and soil environment. Thus, techniques should be developed for the removal and/or recovery of those HMs from point sources in the generating industries. This paper reports our endeavors concerning the development of in-house developed weak cation exchange polyacrylate hydrogel kaolin composites for heavy metals removal. This type of composite enables desirable characteristics and functions including mechanical strength, bed porosity and cost advantages. This paper emphasizes the effect of varying crosslinker (methylenebis(acrylamide)) concentration. The prepared cation exchanger has been subjected to intensive characterization using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray fluorescence (XRF) and Brunauer Emmett and Teller (BET) method. Moreover, the performance was investigated using synthetic and real wastewater for an industrial complex east of Cairo. Simulated and real wastewater compositions addressed; Cr, Co, Ni, and Pb are in the range of (92-115), (91-103), (86-88) and (99-125), respectively. Adsorption experiments have been conducted in both batch and column modes. In general, batch tests revealed enhanced cation exchange capacities of 70, 72, 78.2 and 99.9 mg/g from single synthetic wastes while, removal efficiencies of 82.2, 86.4, 44.4 and 96% were obtained for Cr, Co, Ni and Pb, respectively from mixed synthetic wastes. It is concluded that the mixed synthetic and real wastewaters have lower adsorption capacities than single solutions. It is worth mentioned that Pb attained higher adsorption capacities with comparable results in all tested concentrations of synthetic and real wastewaters. Pilot scale experiments were also conducted for mixed synthetic waste in a fluidized bed column for 48 hour cycle time which revealed 86.4%, 58.5%, 66.8% and 96.9% removal efficiency for Cr, Co, Ni, and Pb, respectively with maximum regeneration was also conducted using saline and acid regenerants. Maximum regeneration efficiencies for the column studies higher than the batch ones about by about 30% to 60%. Studies are currently under way to enhance the regeneration efficiency to enable successful scaling up of the adsorption column.

Keywords: polyacrylate hydrogel kaolin, ultrasonic irradiation, heavy metals, adsorption and regeneration

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Authors: Liya Kalinnikova Magnusson

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The present and ongoing research investigation are focusing on special educational origins in Moldova for children with disabilities and its development towards inclusion. The research is coordinated with related research on inclusion in Ukraine and other countries. The research interest in these issues in Moldova is caused by several reasons. The first one is based upon one of the intensive processes of deconstruction of special education institutions in Moldova since 1989. A large number of children with disabilities have been dropping out of these institutions: from 11400 students in 1989 to 5800 students in 1996, corresponding to 1% of all school-age Moldovan learners. Despite the fact that a huge number of students was integrated into regular schools and the dynamics of this data across the country was uneven (the opposite, the dynamics of exclusion was raised in Trans-Dniester on the border of Moldova), the volume of the change was evident and traditional special educational provision was under stable decline. The second reason is tied to transitional challenges, which Moldova met under the force to economic liberalisation that led the country to poverty. Deinstitutionalization of the entire state system took place in the situation of economic polarization of the society. The level of social benefits was dramatically diminished, increasing inequality. The most vulnerable from the comprehensive income consideration were families with many children, children with disabilities, children with health problems, etc.: each third child belonged to the poorest population. In 2000-2001: 87,4% of all families with children had incomes below the minimum wage. The research question raised based upon these considerations has been addressed to the investigation of particular patterns of the origins of special education and its development towards inclusion in Moldova from 1980 until the present date: what is the pattern of special education origins and what are particular arrangements of special education development towards inclusion against inequality? This is a qualitative study, with relevant peer review resources connected to the research question and national documents of educational reforms towards inclusion retrospectively and contemporary, analysed by a content analysis approach. This study utilises long term statistics completed by the respective international agencies as a result of regular monitoring of the implementation of educational reforms. The main findings were composed in three big themes: adoption of the Soviet pattern of special education, ‘endemic stress’ of breaking the pattern, and ‘paradoxes of resolution’.

Keywords: special education, statistics, educational reforms, inclusion, children with disabilities, content analysis

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Authors: Kitti Szoke, Laszlo Szoke, Attila Czompa, Arpad Tosaki, Istvan Lekli

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Autophagy plays an important role in cardiac hypertrophy, which is one of the most common causes of heart failure in the world. This self-degradative catabolic process, responsible for protein quality control, balancing sources of energy at critical times, and elimination of damaged organelles. The autophagic activity can be triggered by starvation, oxidative stress, or pharmacological agents, like rapamycin. This induced autophagy can promote cell survival during starvation or pathological stress. In this study, it is investigated the effect of the induced autophagic process on angiotensin induced hypertrophic H9c2 cells. In our study, it is used H9c2 cells as an in vitro model. To induce hypertrophy, cells were treated with 10000 nM angiotensin-II, and to activate autophagy, 100 nM rapamycin treatment was used. The following groups were formed: 1: control, 2: 10000 nM AT-II, 3: 100 nM rapamycin, 4: 100 nM rapamycin pretreatment then 10000 nM AT-II. The cell viability was examined via MTT (cell proliferation assay) assay. The cells were stained with rhodamine-conjugated phalloidin and DAPI to visualize F-actin filaments and cell nuclei then the cell size alteration was examined in a fluorescence microscope. Furthermore, the expression levels of autophagic and apoptotic proteins such as Beclin-1, p62, LC3B-II, Cleaved Caspase-3 were evaluated by Western blot. MTT assay result suggests that the used pharmaceutical agents in the tested concentrations did not have a toxic effect; however, at group 3, a slight decrement was detected in cell viability. In response to AT-II treatment, a significant increase was detected in the cell size; cells became hypertrophic. However, rapamycin pretreatment slightly reduced the cell size compared to group 2. Western blot results showed that AT-II treatment-induced autophagy, because the increased expression of Beclin-1, p62, LC3B-II were observed. However, due to the incomplete autophagy, the apoptotic Cleaved Caspase-3 expression also increased. Rapamycin pretreatment up-regulated Beclin-1 and LC3B-II, down-regulated p62 and Cleaved Caspase-3, indicating that rapamycin-induced autophagy can restore the normal autophagic flux. Taken together, our results suggest that rapamycin activated autophagy reduces angiotensin-II induced hypertrophy.

Keywords: angiotensin-II, autophagy, H9c2 cell line, hypertrophy, rapamycin

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Authors: Anjali G. Pillai, S. Chadrakaran

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The extensive amount of sludge generated throughout the world, as a part of water treatment works, have caused various social and economic issues, such as a demand on landfill spaces, increase in environmental pollution and raising the waste management cost. With growing social awareness about toxic incinerator emissions and the increasing concern over the disposal of sludge on the agricultural land, the recovery of sewage sludge as a building and construction raw material can be considered as an innovative approach to tackle the sludge disposal problem. The proposed work aims at studying the recycling ability of the sludge, generated from the water treatment process, by incorporating it into the fired clay brick units. The work involves initial study of the geotechnical characteristics of the brick-clay and the sludge. Chemical compatibility of both the materials will be analyzed by X-ray fluorescence technique. The variation in the strength aspects with varying proportions of sludge i.e. 10%, 20%, 30% and 40% in the sludge-clay mix will also be determined by the proctor density test. Based on the optimum moisture content, the sludge-clay bricks will be manufactured in a brick manufacturing plant and the modified brick units will be tested to determine the variation in compressive strength, bulk density, firing shrinkage, shrinkage loss and initial water absorption rate with respect to the conventional clay bricks. The results will be compared with the specifications given in Indian Standards to arrive at the potential use of the new bricks. The durability aspect will be studied by conducting the leachate analysis test using atomic adsorption spectrometry. The lightweight characteristics of the sludge modified bricks will be ascertained with the scanning electron microscope technique which will be indicative of the variation in pore structure with the increase in sludge content within the bricks. The work will determine the suitable proportion of the sludge – clay mix in the brick which can then be effectively implemented. The feasibility aspect of the work will be determined for commercial production of the units. The work involves providing a strategy for conversion of waste to resource. Moreover, it provides an alternative solution to the problem of growing scarcity of brick-clay for the manufacturing of fired clay bricks.

Keywords: eco-bricks, green construction material, sludge amended bricks, sludge disposal, waste management

Procedia PDF Downloads 282

Authors: A. González-Caro, A. M. Merino-Lechuga, D. Suescum-Morales, E. Fernández-Ledesma, J. R. Jiménez, J. M. Fernández-Rodríguez

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Climate change is one of the most significant issues today. Since the 19th century, the rise in temperature has not only been due to natural change, but also to human activities, which have been the main cause of climate change, mainly due to the burning of fossil fuels such as coal, oil and gas. The boom in the construction sector in recent years is also one of the main contributors to CO₂ emissions into the atmosphere; for example, for every tonne of cement produced, 1 tonne of CO₂ is emitted into the atmosphere. Most of the research being carried out in this sector is focused on reducing the large environmental impact generated during the manufacturing process of building materials. In detail, this research focuses on the recovery of waste from olive oil mills. Spain is the world's largest producer of olive oil, and this sector generates a large amount of waste and by-products such as olive pits, “alpechín” or “alpeorujo”. This olive stone by means of a pyrosilisis process gives rise to the production of active carbon. The process causes the carbon to develop many internal spaces. This study is based on the manufacture of porous mortars with Portland cement and natural limestone sand, with an addition of 5% and 10% of activated carbon. Two curing environments were used: i) dry chamber, with a humidity of 65 ± 10% and temperature of 21 ± 2 ºC and an atmospheric CO₂ concentration (approximately 0.04%); ii) accelerated carbonation chamber, with a humidity of 65 ± 10% and temperature of 21 ± 2 ºC and an atmospheric CO₂ concentration of 5%. In addition to eliminating waste from an industry, the aim of this study is to reduce atmospheric CO₂. For this purpose, first, a physicochemical and mineralogical characterisation of all raw materials was carried out, using techniques such as fluorescence and X-ray diffraction. The particle size and specific surface area of the activated carbon were determined. Subsequently, tests were carried out on the hardened mortar, such as thermogravimetric analysis (to determine the percentage of CO₂ capture), as well as mechanical properties, density, porosity, and water absorption. It was concluded that the activated carbon acts as a sink for CO₂, causing it to be trapped inside the voids. This increases CO₂ capture by 300% with the addition of 10% activated carbon at 7 days of curing. There was an increase in compressive strength of 17.5% with the CO₂ chamber after 7 days of curing using 10% activated carbon compared to the dry chamber.

Keywords: olive stone, activated carbon, porous mortar, CO₂ capture, economy circular

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Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

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The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

Procedia PDF Downloads 144

Authors: Narendra Akhadkar, Philippe Delcambre

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In power protection and control products, assembly process variations due to the individual parts manufactured from single or multi-cavity tooling is a major problem. The dimensional and geometrical variations on the individual parts, in the form of manufacturing tolerances and assembly tolerances, are sources of clearance in the kinematic joints, polarization effect in the joints, and tolerance stack-up. All these variations adversely affect the quality of product, functionality, cost, and time-to-market. Variation simulation analysis may be used in the early product design stage to predict such uncertainties. Usually, variations exist in both manufacturing processes and materials. In the tolerance analysis, the effect of the dimensional and geometrical variations of the individual parts on the functional characteristics (conditions) of the final assembled products are studied. A functional characteristic of the product may be affected by a set of interrelated dimensions (functional parameters) that usually form a geometrical closure in a 3D chain. In power protection and control products, the prerequisite is: when a fault occurs in the electrical network, the product must respond quickly to react and break the circuit to clear the fault. Usually, the response time is in milliseconds. Any failure in clearing the fault may result in severe damage to the equipment or network, and human safety is at stake. In this article, we have investigated two important functional characteristics that are associated with the robust performance of the product. It is demonstrated that the experimental data obtained at the Schneider Electric Laboratory prove the very good prediction capabilities of the variation simulation performed using CETOL (tolerance analysis software) in an industrial context. Especially, this study allows design engineers to better understand the critical parts in the product that needs to be manufactured with good, capable tolerances. On the contrary, some parts are not critical for the functional characteristics (conditions) of the product and may lead to some reduction of the manufacturing cost, ensuring robust performance. The capable tolerancing is one of the most important aspects in product and manufacturing process design. In the case of miniature circuit breaker (MCB), the product's quality and its robustness are mainly impacted by two aspects: (1) allocation of design tolerances between the components of a mechanical assembly and (2) manufacturing tolerances in the intermediate machining steps of component fabrication.

Keywords: geometrical variation, product robustness, tolerance analysis, variation simulation

Procedia PDF Downloads 144

Authors: Xining Wang

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After-school educational service has been proven that it could benefit children’s academic performance, socio-emotional skills, and physical health level. However, there is little research demonstrating parents’ perspectives on the choice of after-school educational service from a level of cross-cultural backgrounds. China and Ireland are typical representatives of collectivist countries (e.g., estimated individualism score is 20) and individualist countries (e.g., estimated individualism score is 70) according to Hofstede's cultural dimensions theory. Living in countries with distinguished cultural backgrounds, there is an evident discrepancy in parents’ attitudes towards domestic after-school education and parents’ motivations for choosing after-school educational services. Through conducting a semi-structured interview with 15 parents from China and 15 parents from Ireland, using thematic analysis software (ATLAS) to extract the key information, and applying a comparative approach to process data analysis; results present polarization of Chinese and Irish parents' perspectives and motivations on after-school educational service. For example, Chinese parents tend to view after-school education as a complement to school education. It is a service they purchased for their children to acquire extra knowledge and skills so that they could adapt to the highly competitive educational setting. Given the fact that children’s education is a priority for Chinese families, most parents believe that their children would succeed in the future through massive learning. This attitude reflects that Chinese parents are more likely to apply authoritarian parenting methods and having a strong expectations for their children. Conversely, Irish parents' choice of after-school educational service is a consideration that primarily based on their own situation, secondly, for their family. For instance, with the expansion of the labor market, there is a change in household structure. Irish mothers are more likely to seek working opportunities instead of looking after the family. Irish parents view that after-school educational service is an essential need for themselves and a beneficial component for their family due to the external pressure (e.g., the growing work intensity and extended working hours, increasing numbers of separated families, as well as parents’ pursuit of higher education and promotion). These factors are fundamental agents that encourage Irish parents to choose after-school educational services. To conclude, the findings could provide readers with a better understanding of parents’ disparate and contrasting perspectives on after-school educational services from a multi-culture level.

Keywords: after-school, China, family studies, Ireland, parents

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Authors: Himani Sharma, Amit Agrawal

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Stability of the Cassie and Wenzel wetting states depends on intrinsic contact angle and geometric features on a surface that was exploited in capturing biofluids in microwells. However, the mechanism of imbibition of biofluids in the microwells is not well implied in terms of wettability of a substrate. In this work, we experimentally demonstrated filling dynamics in hydrophilic and hydrophobic microwells by protein solutions. Towards this, we utilized lotus leaf as a mold to fabricate microwells on a Polydimethylsiloxane (PDMS) surface. Lotus leaf containing micrometer-sized blunt-conical shaped pillars with a height of 8-15 µm and diameter of 3-8 µm were transferred on to PDMS. Furthermore, PDMS surface was treated with oxygen plasma to render the hydrophilic nature. A 10µL droplets containing fluorescein isothiocyanate (FITC) - labelled bovine serum albumin (BSA) were rested on both hydrophobic (θa = 108o, where θa is the apparent contact angle) and hydrophilic (θa = 60o) PDMS surfaces. A time-dependent fluorescence microscopy was conducted on these modified PDMS surfaces by recording the fluorescent intensity over a 5 minute period. It was observed that, initially (at t=1 min) FITC-BSA was accumulated on the periphery of both hydrophilic and hydrophobic microwells due to incomplete penetration of liquid-gas meniscus. This deposition of FITC-BSA on periphery of microwell was not changed with time for hydrophobic surfaces, whereas, a complete filling was occurred in hydrophilic microwells (at t=5 mins). This attributes to a gradual movement of three-phase contact line along the vertical surface of the hydrophilic microwells as compared to stable pinning in the hydrophobic microwells as confirmed by Surface Evolver simulations. In addition, if the cavities are presented on hydrophobic surfaces, air bubbles will be trapped inside the cavities once the aqueous solution is placed over these surfaces, resulting in the Cassie-Baxter wetting state. This condition hinders trapping of proteins inside the microwells. Thus, it is necessary to impart hydrophilicity to the microwell surfaces so as to induce the Wenzel state, such that, an entire solution will be fully in contact with the walls of microwells. Imbibition of microwells by protein solutions was analyzed in terms fluorescent intensity versus time. The present work underlines the importance of geometry of microwells and surface wettability of substrate in wetting and effective capturing of solid sub-phases in biofluids.

Keywords: BSA, microwells, surface evolver, wettability

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Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

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G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

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Authors: Donghyun Lee, Cam Nguyen

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Multi-band systems offer a great deal of benefit in modern communication and radar systems. In particular, multi-band antenna-array radar systems with their extended frequency diversity provide numerous advantages in detection, identification, locating and tracking a wide range of targets, including enhanced detection coverage, accurate target location, reduced survey time and cost, increased resolution, improved reliability and target information. An accurate calibration is a critical issue in antenna array systems. The amplitude and phase errors in multi-band and multi-polarization antenna array transceivers result in inaccurate target detection, deteriorated resolution and reduced reliability. Furthermore, the digital beam former without the RF domain phase-shifting is less immune to unfiltered interference signals, which can lead to receiver saturation in array systems. Therefore, implementing integrated front-end architecture, which can support calibration function with low insertion and filtering function from the farthest end of an array transceiver is of great interest. We report a dual K/Ka-band T/R/Calibration switch module with quasi-elliptic dual-bandpass filtering function implementing a Q-enhanced metamaterial transmission line. A unique dual-band frequency response is incorporated in the reception and calibration path of the proposed switch module utilizing the composite right/left-handed meta material transmission line coupled with a Colpitts-style negative generation circuit. The fabricated fully integrated T/R/Calibration switch module in 0.18-μm BiCMOS technology exhibits insertion loss of 4.9-12.3 dB and isolation of more than 45 dB in the reception, transmission and calibration mode of operation. In the reception and calibration mode, the dual-band frequency response centered at 24.5 and 35 GHz exhibits out-of-band rejection of more than 30 dB compared to the pass bands below 10.5 GHz and above 59.5 GHz. The rejection between the pass bands reaches more than 50 dB. In all modes of operation, the IP1-dB is between 4 and 11 dBm. Acknowledgement: This paper was made possible by NPRP grant # 6-241-2-102 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.

Keywords: microwaves, millimeter waves, T/R switch, wireless communications, wireless communications

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Authors: Pedro Cardoso Alves, Ana Lucia Galinkin, José Carlos Ribeiro

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Political participation, can be a somewhat tricky subject to define, not in small part due to the constant changes in the concept fruit of the effort to include new forms of participatory behavior that go beyond traditional institutional channels. With the advent of the internet and mobile technologies, defining political participation has become an even more complicated endeavor, given de amplitude of politicized behaviors that are expressed throughout these mediums, be it in the very organization of social movements, in the propagation of politicized texts, videos and images, or in the micropolitical behaviors that are expressed in daily interaction. In fact, the very frontiers that delimit physical and digital spaces have become ever more diluted due to technological advancements, leading to a hybrid existence that is simultaneously physical and digital, not limited, as it once was, to the temporal limitations of classic communications. Moving away from those institutionalized actions of traditional political behavior, an idea of constant and fluid participation, which occurs in our daily lives through conversations, posts, tweets and other digital forms of expression, is discussed. This discussion focuses on the factors that precede more direct forms of political participation, interpreting the relation between informational habits, ideology, and political efficacy. Though some of the informational habits can be considered political participation, by some authors, a distinction is made to establish a logical flow of behaviors leading to participation, that is, one must gather and process information before acting on it. To reach this objective, a quantitative survey is currently being applied in Brazilian social media, evaluating feelings of political efficacy, social and economic issue-based ideological stances and informational habits pertaining to collection, fact-checking, and diversity of sources and ideological positions present in the participant’s political information network. The measure being used for informational habits relies strongly on a mix of information literacy and political sophistication concepts, bringing a more up-to-date understanding of information and knowledge production and processing in contemporary hybrid (physical-digital) environments. Though data is still being collected, preliminary analysis point towards a strong correlation between information habits and political efficacy, while ideology shows a weaker influence over efficacy. Moreover, social ideology and economic ideology seem to have a strong correlation in the sample, such intermingling between social and economic ideals is generally considered a red flag for political polarization.

Keywords: political efficacy, ideology, information literacy, cyberpolitics

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Authors: Zhen Cao, Yu Zhu, Junxue Fu

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Immunoassay, i.e., antigen-antibody reaction, is crucial for disease diagnostics. To achieve the adequate signal of the antigen protein detection, a large amount of sample and long incubation time is needed. However, the amount of protein is usually small at the early stage, which makes it difficult to detect. Unlike cells and DNAs, no valid chemical method exists for protein amplification. Thus, an alternative way to improve the signal is through particle manipulation techniques to concentrate proteins, among which dielectrophoresis (DEP) is an effective one. DEP is a technique that concentrates particles to the designated region through a force created by the gradient in a non-uniform electric field. Since DEP force is proportional to the cube of particle size and square of electric field gradient, it is relatively easy to capture larger particles such as cells. For smaller ones like proteins, a super high gradient is then required. In this work, three-dimensional Ag/SiO2 nanorods arrays, fabricated by an easy physical vapor deposition technique called as oblique angle deposition, have been integrated with a DEP device and created the field gradient as high as of 2.6×10²⁴ V²/m³. The nanorods based DEP device is able to enrich bovine serum albumin (BSA) protein by 1800-fold and the rate has reached 180-fold/s when only applying 5 V electric potential. Based on the above nanorods integrated DEP platform, an immunoassay of mouse immunoglobulin G (IgG) proteins has been performed. Briefly, specific antibodies are immobilized onto nanorods, then IgG proteins are concentrated and captured, and finally, the signal from fluorescence-labelled antibodies are detected. The limit of detection (LoD) is measured as 275.3 fg/mL (~1.8 fM), which is a 20,000-fold enhancement compared with identical assays performed on blank glass plates. Further, prostate-specific antigen (PSA), which is a cancer biomarker for diagnosis of prostate cancer after radical prostatectomy, is also quantified with a LoD as low as 2.6 pg/mL. The time to signal saturation has been significantly reduced to one minute. In summary, together with an easy nanorod fabrication and integration method, this nanorods based DEP platform has demonstrated highly sensitive immunoassay performance and thus poses great potentials in applications for early point-of-care diagnostics.

Keywords: dielectrophoresis, immunoassay, oblique angle deposition, protein concentration

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